2024-03-29T02:28:49Z
https://soar-ir.repo.nii.ac.jp/oai
oai:soar-ir.repo.nii.ac.jp:00010309
2022-12-14T04:02:04Z
1016:1017
Engineering of Corynebacterium glutamicum with an NADPH-Generating Glycolytic Pathway for L-Lysine Production
Takeno, Seiki
Murata, Ryosuke
Kobayashi, Ryosuke
Mitsuhashi, Satoshi
Ikeda, Masato
A sufficient supply of NADPH is a critical factor in L-lysine production by Corynebacterium glutamicum. Endogenous NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH) of C. glutamicum was replaced with nonphosphorylating NADP-dependent glyceraldehyde 3-phosphate dehydrogenase (GapN) of Streptococcus mutans, which catalyzes the reaction of glyceraldehyde 3-phosphate to 3-phosphoglycerate with the reduction of NADP(+) to NADPH, resulting in the reconstruction of the functional glycolytic pathway. Although the growth of the engineered strain on glucose was significantly retarded, a suppressor mutant with an increased ability to utilize sugars was spontaneously isolated from the engineered strain. The suppressor mutant was characterized by the properties of GapN as well as the nucleotide sequence of the gene, confirming that no change occurred in either the activity or the basic properties of GapN. The suppressor mutant was engineered into an L-lysine-producing strain by plasmid-mediated expression of the desensitized lysC gene, and the performance of the mutant as an L-lysine producer was evaluated. The amounts of L-lysine produced by the suppressor mutant were larger than those produced by the reference strain (which was created by replacement of the preexisting gapN gene in the suppressor mutant with the original gapA gene) by similar to 70% on glucose, similar to 120% on fructose, and similar to 100% on sucrose, indicating that the increased L-lysine production was attributed to GapN. These results demonstrate effective L-lysine production by C. glutamicum with an additional source of NADPH during glycolysis.
Article
APPLIED AND ENVIRONMENTAL MICROBIOLOGY. 76(21): 7154-7160(2010)
AMER SOC MICROBIOLOGY
2010-11
eng
journal article
AM
http://hdl.handle.net/10091/13188
https://soar-ir.repo.nii.ac.jp/records/10309
https://doi.org/10.1128/AEM.01464-10
10.1128/AEM.01464-10
0099-2240
AA00543249
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
76
21
7154
7160
https://soar-ir.repo.nii.ac.jp/record/10309/files/Engineering_Corynebacterium_glutamicum.pdf
application/pdf
497.3 kB
2015-09-25